A virtual workshop for precise HDR-mediated genome engineering with CRISPR-Cas9
The use of CRISPR-Cas9 to create targeted double-strand breaks in genomic DNA has greatly simplified strategies for precise gene editing using the homology-directed repair (HDR) mechanism. This approach enables knock-in of fluorescent reporters, SNP repair, exact deletions, and many other genomic alterations. However, designing target-specific guide RNAs and DNA donor templates for achieving robust cleavage and repair has many challenges. In addition, the efficiency of HDR-mediated repair is often low. In this workshop, we will demonstrate the use of free, intuitive, online tools to design and order the necessary reagents to overcome these obstacles with HDR-mediated gene editing applications.
Learning Objectives:
- Considerations for designing ideal guide RNAs and DNA oligo or plasmid repair templates for HDR-mediated CRISPR-Cas9 applications
- Use of online tools for guide RNA and DNA repair templates for HDR-mediated gene editing applications
- Optimization of experimental conditions to achieve robust gene editing
- Demonstration of successful HDR-mediated CRISPR Cas9 experiments
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